The discovery of Hsp70 domain with cell-penetrating activity

Elena Y. Komarova, Darya A. Meshalkina, Nikolay D. Aksenov, Ivan M. Pchelin, Elena Martynova, Boris A. Margulis, Irina V. Guzhova

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

Chaperone Hsp70 can cross the plasma membrane of living cells using mechanisms that so far have not received much research attention. Searching the part of the molecule that is responsible for transport ability of Hsp70, we found a cationic sequence composed of 20 amino acid residues on its surface, KST peptide, which was used in further experiments. We showed that KST peptide enters living cells of various origins with the same efficiency as the full-length chaperone. KST peptide is capable of carrying cargo with a molecular weight 30 times greater than its own into cells. When we compared the membrane-crossing activity of KST peptide in complex with Avidin (KST–Av complex) with that of similarly linked canonical TAT peptide, we found that TAT peptide penetrated SK-N-SH human neuroblastoma cells at a similar rate and efficiency as the KST peptide. Furthermore, KST peptide can carry protein complexes consisting of a specific antibody coupled to the peptide through the Avidin bridge. An antibody to Hsp70 delivered to SK-N-SH cells with high expression level of Hsp70 reduced the protective power of the chaperone and sensitized the cells to the pro-apoptotic effect of staurosporine. We studied the mechanisms of penetration of KST–Av and full-length Hsp70 inside human neuroblastoma SK-N-SH and human erythroleukemia K-562 cells and found that both used an active intracellular transport mechanism that included vesicular structures and negatively charged lipid membrane domains. Competition analysis of intracellular transport showed that the chaperone reduced intracellular penetration of KST peptide and conversely KST peptide prevented Hsp70 transport in a dose-dependent manner.

Original languageEnglish (US)
Pages (from-to)343-354
Number of pages12
JournalCell Stress and Chaperones
Volume20
Issue number2
DOIs
StatePublished - Feb 13 2015

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Peptides
Avidin
Neuroblastoma
Cells
Leukemia, Erythroblastic, Acute
Staurosporine
Antibodies
Active Biological Transport
Cell membranes
Membrane Lipids
Molecular Weight
Molecular weight
Cell Membrane
Membranes
Amino Acids
Molecules
Research
Proteins
Experiments

Keywords

  • Cell-penetrating peptides
  • Hsp70
  • Intracellular transport
  • Membrane
  • Vesicles

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology

Cite this

Komarova, E. Y., Meshalkina, D. A., Aksenov, N. D., Pchelin, I. M., Martynova, E., Margulis, B. A., & Guzhova, I. V. (2015). The discovery of Hsp70 domain with cell-penetrating activity. Cell Stress and Chaperones, 20(2), 343-354. https://doi.org/10.1007/s12192-014-0554-z

The discovery of Hsp70 domain with cell-penetrating activity. / Komarova, Elena Y.; Meshalkina, Darya A.; Aksenov, Nikolay D.; Pchelin, Ivan M.; Martynova, Elena; Margulis, Boris A.; Guzhova, Irina V.

In: Cell Stress and Chaperones, Vol. 20, No. 2, 13.02.2015, p. 343-354.

Research output: Contribution to journalArticle

Komarova, EY, Meshalkina, DA, Aksenov, ND, Pchelin, IM, Martynova, E, Margulis, BA & Guzhova, IV 2015, 'The discovery of Hsp70 domain with cell-penetrating activity', Cell Stress and Chaperones, vol. 20, no. 2, pp. 343-354. https://doi.org/10.1007/s12192-014-0554-z
Komarova EY, Meshalkina DA, Aksenov ND, Pchelin IM, Martynova E, Margulis BA et al. The discovery of Hsp70 domain with cell-penetrating activity. Cell Stress and Chaperones. 2015 Feb 13;20(2):343-354. https://doi.org/10.1007/s12192-014-0554-z
Komarova, Elena Y. ; Meshalkina, Darya A. ; Aksenov, Nikolay D. ; Pchelin, Ivan M. ; Martynova, Elena ; Margulis, Boris A. ; Guzhova, Irina V. / The discovery of Hsp70 domain with cell-penetrating activity. In: Cell Stress and Chaperones. 2015 ; Vol. 20, No. 2. pp. 343-354.
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